摘要
目的探讨应用α1（I）型前胶原基因反义寡聚脱氧核苷酸对增生性瘢痕成纤维细胞生长、胶原合成等生物学特性的影响。方法根据转染液分3组，实验组为脂质体加α1（I）型前胶原基因反义寡聚脱氧核苷酸；对照组1为脂质体组；对照组2为培养基空白对照组。用脂质体法将α1（I）型前胶原基因反义寡聚脱氧核苷酸转染增生陛髓魔成纤维细胞并运细胞计数法绘制细胞生长曲线；电子透射电镜观察转染后各组ttF超微结构的变化；转染后0．5、1、3、5、7d分别提取各组细胞总RNA，行RT-PCR后测定各组I胶原mRNA表达量；Western．bloring法检测转染后各组琏!胶原蛋白的变化。结果①反义寡聚脱氧核苷酸对增生陛瘢痕成纤维细胞生长增殖无抑制；②实验组增生l生瘢痕成纤维细胞蛋白合成的细胞器呈非活跃状态；③与对照组比较，实验组HF的a。（I）型前胶原蛋白mRNA的表达和例胶原合成量明显减少。结论α1（I）型前胶原基因反义寡聚脱氧核苷酸能抑制人增生性瘢痕成纤维细胞的α1（I）型前胶原蛋白mRNA的表达和理I胶原蛋白的合成，提示可能抑制瘢痕增生。
Objective To study the effect of antisense oligodeoxynucleotides of type oq （I） precollagen gene on growth and the collagen synthesis of human hypertrophic scar fibroblasts. Methods The HF were divided into 3 groups according to the transfection liquids ： the experimental group with LipofectAMINETM and type cq （I） prec- ollagen gene antisense oligodeoxynucleotides, the control group 1 with only LipofectAMINETM and the control group 2 with only DMEM culture medium as the control group. At 0.5, 1, 3, 5 and 7 days after transfection, different measures were adopted to evaluate the biological characteristics of hypertrophic scar fibroblasts, such as the fibroblast growth curve protracted by the cell count technique, the ultra-structure observed by transmission eletron microscopy （TEM）, total RNA of the hypertrophic scar fibroblasts were extracted and the expression level of type α1 （I） precal- lagen mRNA was detected by RT-PCR,the type I collagen synthesis of hypertrophic scar fibroblasts was tested by the westem-bloting and compared with the control group. Results The fibroblast proliferation was not inhibited by antisense aligodeoxynucleotides. The organelles for the protein synthesis in the experimental group was not in ac- tive state. Compared with the control group, the expression level of type α （I） precollege mRNA and protein syn- thesis of hypertrophic scar fibroblasts in the experimental group were obviously decreased. Conclusion The anti- sense aligodeoxynucleotides inhibits availably the expression level of type α（I） precollagen mRNA and the protein synthesis of I collagen of hypertrophic scar fibrobtasts cultured in vitro, which can inhibint the scar hyperplasia.
出处
《中国美容整形外科杂志》
CAS
2013年第3期152-156,共5页
Chinese Journal of Aesthetic and Plastic Surgery
基金
广东省科委公关项目（9827817）
国家自然科学基金面上项目（81071557）
